LOCUS       sequence-429135-        6697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human CALR wt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6697)
  TITLE     Masubuchi et al., 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6697)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1/CALR wt
FEATURES             Location/Qualifiers
     source          1..6697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      34..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(121..140)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     331..352
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(471..491)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        476..805
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      656..791
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     718..737
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             872..1666
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(926..945)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1536..1555
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1840..1961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1877..1896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1931..1950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2010..2026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2010..2026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2023..2045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2034..2050
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2058..2064,2065..2082,2083..2088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2103..2124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2241..2258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2412..3000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2492..2511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3171..3962,3963..4031))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3794..3813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4032..4136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4211..4230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4402..4781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4782..4985
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4936..4956
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5030..5049
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5030..5048
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6455..6472)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6461..6685
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
       61 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
      121 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      181 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
      241 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      301 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      361 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
      421 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
      481 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
      541 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      601 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
      661 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
      721 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
      781 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
      841 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
      901 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
      961 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     1021 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     1081 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     1141 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     1201 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     1261 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     1321 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     1381 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     1441 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     1501 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     1561 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     1621 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     1681 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     1741 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     1801 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     1861 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1921 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     1981 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2041 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2101 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2161 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2221 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2281 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2341 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2401 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2461 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2521 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2581 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2641 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2701 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2761 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2821 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2881 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2941 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3001 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3061 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     4201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     4261 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     4321 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     4381 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     4441 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     4501 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     4561 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     4621 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     4681 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     4741 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     4801 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     4861 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     4921 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     4981 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     5041 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
     5101 cactagtcca gtgtggtgga attcgccacc atgctgctat ccgtgccgct gctgctcggc
     5161 ctcctcggcc tggccgtcgc cgagcctgcc gtctacttca aggagcagtt tctggacgga
     5221 gacgggtgga cttcccgctg gatcgaatcc aaacacaagt cagattttgg caaattcgtt
     5281 ctcagttccg gcaagttcta cggtgacgag gagaaagata aaggtttgca gacaagccag
     5341 gatgcacgct tttatgctct gtcggccagt ttcgagcctt tcagcaacaa aggccagacg
     5401 ctggtggtgc agttcacggt gaaacatgag cagaacatcg actgtggggg cggctatgtg
     5461 aagctgtttc ctaatagttt ggaccagaca gacatgcacg gagactcaga atacaacatc
     5521 atgtttggtc ccgacatctg tggccctggc accaagaagg ttcatgtcat cttcaactac
     5581 aagggcaaga acgtgctgat caacaaggac atccgttgca aggatgatga gtttacacac
     5641 ctgtacacac tgattgtgcg gccagacaac acctatgagg tgaagattga caacagccag
     5701 gtggagtccg gctccttgga agacgattgg gacttcctgc cacccaagaa gataaaggat
     5761 cctgatgctt caaaaccgga agactgggat gagcgggcca agatcgatga tcccacagac
     5821 tccaagcctg aggactggga caagcccgag catatccctg accctgatgc taagaagccc
     5881 gaggactggg atgaagagat ggacggagag tgggaacccc cagtgattca gaaccctgag
     5941 tacaagggtg agtggaagcc ccggcagatc gacaacccag attacaaggg cacttggatc
     6001 cacccagaaa ttgacaaccc cgagtattct cccgatccca gtatctatgc ctatgataac
     6061 tttggcgtgc tgggcctgga cctctggcag gtcaagtctg gcaccatctt tgacaacttc
     6121 ctcatcacca acgatgaggc atacgctgag gagtttggca acgagacgtg gggcgtaaca
     6181 aaggcagcag agaaacaaat gaaggacaaa caggacgagg agcagaggct taaggaggag
     6241 gaagaagaca agaaacgcaa agaggaggag gaggcagagg acaaggagga tgatgaggac
     6301 aaagatgagg atgaggagga tgaggaggac aaggaggaag atgaggagga agatgtcccg
     6361 ggccaggcca aggacgagct gtaggaattc tgcagatatc cagcacagtg gcggccgctc
     6421 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     6481 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     6541 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     6601 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     6661 gctggggatg cggtgggctc tatggcttct gaggcgg
//
